The Study Of Dandruff Efficacy Of Immortals Pill

ObjectiveThe purpose of this study is to detect Immortals Pill suppress Malassezia and it on the role of LPS induced inhibition of the Immortalized human Keratinocytes (HaCaT). Provide efficacy experimental evidence to the anti dandruff hair care products which contains traditional Chinese medicine ingredients.MethodsFirst, using a10-fold amount of60%ethanol reflux extraction2hours for Desefruit Pittany Poot-baik, Cacumen Platycladi, Lightyellow Sothora Root, Radix Angelicae Sinendsis, repeat3times, then centrifugal and filter, recovery of ethanol made from liquid extract, finally, diluted with culture medium to appropriate concentrations of liquid. The experimental group was divided into three groups, Traditional Chinese Medicine group:Desefruit Pittany Poot-baik, Cacumen Platycladi, Lightyellow Sothora Root, Radix Angelicae Sinendsis, Immortals Pill (Desefruit Pittany Poot-baik+Cacumen Platycladi), Immortals Pill+Lightyellow Sothora Root, Immortals Pill+Radix Angelicae Sinendsis, and Four Chineseherbal medicine mixture (Immortall pill+Radix Angelicae Sinendsis+Cacumen Platycladi); Monomer group:Matrine, Oxymatrine, Dictamnine, Ferulic acid, Quercetin; Positive control group: Zinc Pyrithione (ZPT).Second, using the method of Liquid drugs medium, cultured (0.5～1.0)×108CFU/mL Malassezia globosa, Malassezia Furfur, Malassezia Sympodialisby and Malassezia Obtuse by drug-containing medium (The final concentration of each group:Traditional Chinese Medicine group is0.977mg/mL～1000mg/m L; Monomer group is4.883×10-4mg/mL-0.5mg/mL; ZPT is2.441×10-2μg/mL～25μg/mL). Compared with growth control group, The minimum inhibitory conc entration (MIC) is that the minimum liquid concentration of Malassezia w as inhibited80%.Finally, Using the method of Methyl Thiazolyl Tetrazolium (MTT) and Trypan blue (TB), cultured HaCaT by drug-containing medium (The final co ncentration of each group:Traditional Chinese Medicine group is20mg/mL,2mg/mL,0.2mg/mL; Monomer group is5×10-2mg/mL,5×10-3mg/mL,5×10-4mg/mL; ZPT is0.2%、2×10-2%、2×10-3%). Detect the Cell viability (CV) of HaCaT.ResultsThe minimum Inhibitive Concentration (MIC) or The Relative minimum Inhibitive Concentration (RMIC) sorted from small to large were ZPT (MIC:1.2×10-3mg/mL), Dictamnine (MIC:0.2mg/mL, RMIC:43.2mg/mL), Matrine (MIC:0.5mg/mL, RMIC:74.5mg/mL), lightyellow Sothora Root (MIC:114.9mg/mL), Cacumen Platycladi (MIC:119. lmg/mL), Oxymatrine (MIC:lmg/mL, RMIC:171.5mg/mL), Immortals pill (182.4mg/mL), Four Chinese herbal medicine mixture (MIC:193.7mg/mL), Desefruit Pittany Poot-baik (MIC:240.3mg/mL), Immortals Pill+lightyellow Sothora Root (MIC:242.2mg/mL), Immortals Pill+Desefruit Pittany Poot-baik Root (MIC:248.7mg/mL), Quercetin (MIC:319. Omg/mL), Radix Angelicae Sinensis (MIC:0.4mg/mL, RMIC:391.4mg/mL), Desefruit Pittany Poot-baik (MIC:768.8mg/mL) and Ferulic acid (MIC:0.5mg/mL, RMIC:1042.2mg/mL). The MIC or RMIC of group Radix Desefruit Pittany Poot-baik, Ferulic acid and Angelicae Sinensis are larger then other groups, and the difference was statistically significant.Groups effective to suppress the Inflammation of HaCaT in both MTT and TB assay are:Immortals Pill (2mg/mL), Angelicae Sinensis (0.5mg/mL), Ferulic acid (0.5mg/mL), Dictamnine (0.5mg/mL). Groups were measured the toxic side effects to HaCaT in MTT assay:3toxicity:ZPT (0.2%), ZPT (2%); Groups were measured the toxic side effects to HaCaT in TB assay:5toxicity:ZPT (2%), Cacumen Platycladi (200mg/mL,20mg/mL), Four Chinese herbal medicine mixture (200mg/mL), Lightyellow Sothora Root (200mg/mL);3toxicity:Immortals Pill (200mg/mL,20mg/mL), Four Chinese herbal medicine mixture (20mg/mL). Groups were measured the toxic side effects to HaCaT in both MTT and TB assay:ZPT (0.2%). ZPT (2%). Other Traditional Chinese Medicine group and Monomer group were not detected in HaCaT cytotoxic.Conclusion①Immortals Pill and ZPT both can effectively inhibit the growth of Malassezia. cure dandruff. But Immortals Pill is safety than ZPT;②In Traditional Chinese Medicine group, the MIC of Immortals Pill is smallest. Just Immortals Pill can suppress the inflammation of HaCaT effectively in both MTT and TB assay. Immortals pill is more effectively to inhibit dandruff than other traditional chinese medicine groups in this study;③In Monomer group, when Ferulic acid and Matrine are0.5mg/mL, they can inhibit Malassezia and suppress the inflammation of HaCaT effectively. They both non-toxic to HaCaT. Matrine is more effective than Immortals Pill to inhibit the Malassezia but Imortals Pill is more effective than Matrine to suppress the inflammation of HaCaT; Ferulic acid is less effective than Immortals Pill both in the inhibition of Malassezia and the suppression of HaCaT’s inflammation.